title: Presence of Epstein-Barr virus latency type III at the single cell level in post-transplantation lymphoproliferative disorders and AIDS related lymphomas.
abstract: OBJECTIVE
  To investigate the expression pattern of Epstein-Barr virus (EBV) latent genes at the single cell level in post-transplantation lymphoproliferative disorders and acquired immunodefiency syndrome (AIDS) related lymphomas, in relation to cellular morphology.
  
  
  METHODS
  Nine post-transplantation lymphoproliferative disorders and three AIDS related lymphomas were subjected to immunohistochemistry using monoclonal antibodies specific for EBV nuclear antigen 1 (EBNA1) (2H4), EBNA2 (PE2 and the new rat anti-EBNA2 monoclonal antibodies 1E6, R3, and 3E9), and LMP1 (CS1-4 and S12). Double staining was performed combining R3 or 3E9 with S12.
  
  
  RESULTS
  R3 and 3E9 anti-EBNA2 monoclonal antibodies were more sensitive than PE2, enabling the detection of more EBNA2 positive lymphoma cells. Both in post-transplantation lymphoproliferative disorders and AIDS related lymphomas, different expression patterns were detected at the single cell level. Smaller neoplastic cells were positive for EBNA2 but negative for LMP1. Larger and more blastic neoplastic cells, sometimes resembling Reed-Sternberg cells, were LMP1 positive but EBNA2 negative (EBV latency type II). Morphologically intermediate neoplastic cells coexpressing EBNA2 and LMP1 (EBV latency type III), were detected using R3 and 3E9, and formed a considerable part of the neoplastic population in four of nine post-transplantation lymphoproliferative disorders and two of three AIDS related lymphomas. All samples contained a subpopulation of small tumour cells positive exclusively for Epstein-Barr early RNA and EBNA1. The relation between cellular morphology and EBV expression patterns in this study was less pronounced in AIDS related lymphomas than in post-transplantation lymphoproliferative disorders, because the AIDS related lymphomas were less polymorphic than the post-transplantation lymphoproliferative disorders.
  
  
  CONCLUSIONS
  In post-transplantation lymphoproliferative disorders and AIDS related lymphomas, EBV latency type III can be detected by immunohistochemistry in a subpopulation of tumour cells using sensitive monoclonal antibodies R3 and 3E9. Our data suggest that EBV infected tumour cells in these lymphomas undergo gradual changes in the expression of EBV latent genes, and that these changes are associated with changes in cellular morphology.
authors.affiliation: Department of Pathology, University Hospital Vrije Universiteit, Amsterdam, Netherlands.
authors.forename: A A
authors.initials: AA
authors.lastname: Brink
authors.forename: D F
authors.initials: DF
authors.lastname: Dukers
authors.forename: A J
authors.initials: AJ
authors.lastname: van den Brule
authors.forename: J J
authors.initials: JJ
authors.lastname: Oudejans
authors.forename: J M
authors.initials: JM
authors.lastname: Middeldorp
authors.forename: C J
authors.initials: CJ
authors.lastname: Meijer
authors.forename: M
authors.initials: M
authors.lastname: Jiwa
mesh_terms: D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D016026:Bone Marrow Transplantation / Q000276:immunology; D002460:Cell Line; D019309:Epstein-Barr Virus Nuclear Antigens / Q000032:analysis*; D004854:Herpesvirus 4, Human / Q000302:isolation & purification* / Q000502:physiology; D006801:Humans; D016867:Immunocompromised Host; D007124:Immunoenzyme Techniques; D016030:Kidney Transplantation / Q000276:immunology; D016483:Lymphoma, AIDS-Related / Q000821:virology*; D008232:Lymphoproliferative Disorders / Q000276:immunology / Q000821:virology*; D014763:Viral Matrix Proteins / Q000032:analysis; D017735:Virus Latency
chemical_list: D000911:Antibodies, Monoclonal; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D019309:Epstein-Barr Virus Nuclear Antigens; D014763:Viral Matrix Proteins